Time-lapse Monitoring System Research Articles

Obstetric and perinatal outcomes of pregnancies conceived with embryos cultured in a time-lapse monitoring system

To compare obstetric and perinatal outcomes of singleton pregnancies resulting from embryos incubated in a time-lapse system (TLS) with those of embryos grown in standard IVF incubators (SI). Retrospective description of a cohort of patients who conceived during a randomized, controlled trial. Private university-affiliated IVF center. Of 856 randomized patients, 378 gave birth to a live-born infant: 216 of the deliveries originated from embryos incubated in TLS, and 162 deliveries were from embryos cultured in SI. Embryo incubation and selection in TLS. Delivery and neonatal outcomes. No significant differences were observed in the baseline characteristics of the study population. The delivery rate was 49.3% (TLS) vs. 40.0% (SI), and multiple deliveries were higher in the TLS group: 31.0% (67 of 216) vs. 24.7% (40 of 162) in the SI group. When singleton pregnancies were analyzed no differences were found between the two groups in the rate of obstetric problems with respect to weeks at delivery: 38.8 (95% confidence interval [CI] 38.4-39.1) (TLS) vs. 39.5 (95% CI 38.0-39.9) (SI); preterm births (<37weeks): 10.7% (TLS) vs. 12.3% (SI); and very preterm births (<34weeks): 2.9% (TLS) vs. 3.3% (SI). No statistical differences were found in neonatal outcomes such as birth weight: 3,163g (95% CI 3,035-3,292g) (TLS) vs. 3,074 (95% CI 2,913-3,236) (SI); low birth weight (<2,500g): 12.8% (TLS) vs. 12.3% (SI); very low birth weight (<1,500g): 2.0% (TLS) vs. 2.4% (SI); or height: 50.3cm (95% CI 49.6-50.9cm) (TLS) vs. 49.7 (95% CI 48.9-50.4cm) (SI). No major malformations or perinatal mortality were found in either of the two groups. No detrimental effects were observed in obstetric and perinatal outcomes when a time-lapse incubator was used rather than a more widely used conventional incubator. As far as we know this is the first report from a randomized study of the neonatal outcomes of time-lapse monitoring. Our results suggest that this technology is an effective and safe alternative for embryo incubation, though trials of larger numbers of patients are required to further confirm our conclusions. NCT01549262.

Morphokinetic parameters from a time-lapse monitoring system cannot accurately predict the ploidy of embryos.

This study aimed to test whether there is an association between embryo morphokinetic parameters and ploidy status. Patients with high risk of aneuploidy were analyzed by time-lapse microscopy combined with preimplantation genetic screening (PGS). Accordingly, 256 blastocysts from 75 patients were subjected to trophectoderm biopsy and microarray comparative genomic hybridization (array-CGH). Blastocyst development process was analyzed using time-lapse images. Morphokinetic parameters: tPNf, t2, t3, t4, t5, t8, t9, tcom, tM, tSB, tB, tEB, CC1, CC2, CC3, S2, S3, t5-t2, and tB-tSB showed no significant difference in euploid embryos compared to aneuploid counterparts. In addition, two risk models based on previously published morphokinetic parameters failed to segregate euploid from aneuploid embryos. Morphokinetic parameters subjected to investigation in the present study failed to improve the chance of selecting euploid embryos.

Investigation of the Developmental Potential and Developmental Kinetics of Bovine Parthenogenetic and Somatic Cell Nuclear Transfer Embryos Using a Time-Lapse Monitoring System.

A time-lapse monitoring system has predictive value for selecting good-quality embryos with the highest implantation potential. Using this new tool, we investigated the developmental potential and developmental kinetics of bovine parthenogenetic (PA) and two types of somatic cell nuclear transfer (NT) embryos. Bovine non-transgenic ear cells (bECs) or transgenic cells (bTGCs) were used as donor cells. The cleavage and blastocyst development rates did not significantly differ among the PA, NT-bEC, and NT-bTGC groups, and first cleavage occurred an average of 19.3 hours (n = 70), 21.6 hours (n = 60), and 21.3 hours (n = 62) after activation, respectively (20.4 hours [n = 192] for all embryos). When embryos were classified into early cleaving (≤20 hours) and late cleaving (>20 hours) groups, the blastocyst formation rate was much higher in the early cleaving groups (PA, 46%; NT-bEC, 50%; NT-bTGC, 39%) than in the late cleaving groups (PA, 18%; NT-bEC, 23%; NT-bTGC, 28%), while the percentage of embryos whose development was blocked between the two- and eight-cell stages was increased in the late cleaving groups. The percentage of embryos classified as early cleaving with a normal morphology was twofold higher in the PA group (20.0%, n = 14) than in the NT-bTGC group (9.7%, n = 6). The timing of each developmental stage varied widely; the timing of first cleavage varied from 7.6 hours in the PA group to 34.5 hours in the NT-bEC group and the timing of expanded/hatching blastocyst appearance varied from 141.6 hours in the PA group to 196.3 hours in the NT-bTGC group, differences of 26.9 and 54.7 hours, respectively (PA>NT-bEC>NT-bTGC). These results demonstrate that time-lapse monitoring provides novel data regarding individual embryo developmental kinetics and helps to predict developmental potential for improved bovine NT embryo selection based on early cleavage (≤20 hours) and normal morphology.

Embryo culture using a time-lapse monitoring system improves live birth rates compared with a conventional culture system: a prospective cohort study

In this prospective cohort study, the effects of a time-lapse monitoring system on embryo quality and clinical pregnancy outcomes were assessed. A total of 608 patients undergoing in vitro fertilization between April 2013 and June 2014 at our institution were recruited for this study and group-matched into a time-lapse monitoring (TLM) (N = 304) or a standard incubator (SI) (N = 304). The patients’ characteristics in the TLM and SI groups were not significantly different. The TLM group showed a significantly higher transferable embryo ratio at Day 3 (61.65% vs. 52.87%; p < 0.0010, RR =1.10 [1.02, 1.19]), a higher number of transferable embryos (4.71 ± 2.38 vs. 4.09 ± 2.35; p = 0.0053, SMD =0.26 [0.06, 0.46]) and number of good-quality embryos cryopreserved at Day 3 (2.72 ± 2.35 vs. 2.11 ± 2.33; p = 0.0056, SMD =0.26 [0.06, 0.46]). In addition, the implantation and clinical pregnancy rates were not statistically significant between the TLM and SI groups. However, the TLM group had a higher ongoing pregnancy rate (67.32% vs. 57.22%; p = 0.0410) and live birth rate (65.37% vs. 55%; p = 0.0380) compared with the SI group. The observed beneficial effects could be the result of a more stable environment provided by the TLM system.

Comparison of the morphokinetic parameters of embryos according to ovarian reserve in IVF cycles

The aim of this study is to evaluate the impact of ovarian reserve and age of women on early morphokinetic parameters of embryos with a time-lapse monitoring system. In total, 197 infertile couples with poor ovarian reserve (Group 1, n = 41), normal ovarian reserve (Group 2, n = 59), or polycystic ovaries (Group 3, n = 97) were included. The time from insemination to the following events were analyzed: pronuclear fading (Pnf) and cleavage to 2, 3, 4 and 5 cells. The optimal ranges for morphokinetic parameters of t5, s2 and cc2 in each group were also evaluated. In total, 1144 embryos were evaluated. Morphokinetic parameters did not differ statistically between the groups. Data were analyzed according to different age groups (20–30, 30–40, >40). The morphokinetic parameters did not differ statistically in Group 1 and 3. In Group 2, the times from insemination to tPnf, t2, t3, t4 were significantly shorter in the younger age group than the older age group (p < 0.05). The percentages of optimal embryos, according to t5, s2 and cc2, did not differ statistically between the groups. In conclusion, ovarian reserve did not seem to affect the morphokinetic parameters of embryos.

Near-infrared laser irradiation improves the development of mouse pre-implantation embryos

The aim of the present study was to assess the effects of near-infrared laser irradiation on the in vitro development of mouse embryos. Female ICR mice were superovulated with pregnant mare serum gonadotropin and human chorionic gonadotropin (hCG), and mated with male mice. Two-cell stage embryos were collected 40 h after administering hCG and cultured in M16 medium. Two-cell embryos (0 h after culture), 8-cell embryos (approx. 30 h after culture), morula (approx. 48 h after culture), and blastocysts (approx. 73 h after culture) were irradiated at 904 nm for 60 s. These embryos were cultured in a time-lapse monitoring system and the timing of blastocyst hatching was evaluated. Some of the irradiated blastocysts were transferred to the uterine horns of pseudopregnant recipients immediately after irradiation. Pregnancy rates, and offspring growth and fertility, were evaluated. Near-infrared laser irradiation increased the speed of in vitro mouse embryo development. In irradiated blastocysts, hatching was faster than in control (non-irradiated) blastocysts (18.4 vs. 28.2 h, P < 0.05). When 195 irradiated blastocysts were transferred to 18 pseudopregnant mice, all became pregnant and 92 (47.2%) normal-looking pups were born alive. When 182 control blastocysts were transferred to 17 pseudopregnant mice, 14 (82.4%) became pregnant and 54 (29.7%) normal-looking pups were born alive. The growth trajectories (up to 5 weeks) of offspring from irradiated blastocysts were similar to those from control blastocysts. Second generation offspring from transplanted animals were all fertile. These results indicate that near-infrared laser irradiation improves the quality of mouse embryo development in vitro, and increases the live birth rate without affecting the normality of the offspring. Thus, the near-infrared laser method may enhance the quality of embryos and contribute to improvements in reproductive technologies in mammals.

Blastomere biopsy for PGD delays embryo compaction and blastulation: a time-lapse microscopic analysis.

The purpose of the study was to explore the effect of blastomere biopsy for preimplantation genetic diagnosis (PGD) on the embryos' dynamics, further cleavage, development, and implantation. The study group included 366 embryos from all PGD treatments (September 2012 to June 2014) cultured in the EmbryoScope™ time-lapse monitoring system. The control group included all intracytoplasmic sperm injection (ICSI) embryos cultured in EmbryoScope™ until day 5 during the same time period (385 embryos). Time points of key embryonic events were analyzed with an EmbryoViewer™. Most (88%) of the embryos were biopsied at ≥8 cells. These results summarize the further dynamic development of the largest cohort of biopsied embryos and demonstrate that blastomere biopsy of cleavage-stage embryos significantly delayed compaction and blastulation compared to the control non-biopsied embryos. This delay in preimplanation developmental events also affected postimplantation development as observed when the dynamics of non-implanted embryos (known implantation data (KID) negative) were compared to those of implanted embryos (KID positive). Analysis of morphokinetic parameters enabled us to explore how blastomere biopsy interferes with the dynamic sequence of developmental events. Our results show that biopsy delays the compaction and the blastulation of the embryos, leading to a decrease in implantation.

The morphokinetic characteristics of embryos derived from pcos patients

Polycystic ovary syndrome (PCOS) patients are extremely sensitive to stimulation with exogenous gonadotropins and are at an increased risk of developing ovarian hyperstimulation syndrome (OHSS) during ART treatment. Such patients are therefore likely to greatly benefit from using the in vitro maturation (IVM) technique. However, IVM oocytes have been found to exhibit lower viability and developmental potential than in vivo matured oocytes. The objective of this study was to compare morphokinetics of early stage embryos from IVM oocytes to that of mature oocytes obtained after controlled ovarian stimulation (COS) in PCOS patients, while mature oocytes from stimulated non-PCOS ovaries served as a control. Retrospective study. From Jan. 2014 to Dec. 2015, zygotes were divided into three groups according to oocyte maturity and patient characteristics. A total of 66 zygotes derived from 15 IVM cycles in PCOS patients (Group A), 33 zygotes from 8 COS cycles in PCOS (Group B), and 43 zygotes from 10 COS cycles in non-PCOS patients (Group C) were involved. Following ICSI insemination, morphokinetic parameters of each zygote were recorded and evaluated by a time-lapse monitoring system (EmbryoScopeTM). In Group B, embryos developing to the 3-cell and 4-cell stage were significantly slower than those in Group C (39.2 ± 6.6 hours vs. 35.1 ± 67.2 hours, 43.2 ± 7.5 hours vs. 39.5 ± 6.7 hours) (P <0.05). The time for the initiation of the compaction and the time to the morula stage were remarkably shorter in Group A (75.5 ± 13.6 hours, 82.9 ± 8.5 hours) and Group B (74.6 ± 17.6 hours, 83.4 ± 18.9 hours) than in Group C (84.2 ± 7.5 hours, 96.8 ± 8.0 hours) (P <0.05). The time for the initiation of blastulation and the time to full blastocyst stage were significantly shorter in Group A (92.8 ± 7.4 hours, 104.3 ± 9.5 hours) than in Group C (102.6 ± 5.7 hours, 114.4 ± 7.0 hours) (P <0.05). Embryos derived from PCOS patients, regardless of the maturity at oocyte retrieval, showed faster development from the initiation of compaction onwards in comparison to non-PCOS patients. These results suggest that the state of PCOS may cause changes in morphokinetic behavior of early embryos.

Time-lapse embryo morphokinetics following GnRH agonist or hCG triggering

GnRH agonist triggering is used instead of hCG in antagonist cycles, in order to diminish ovarian hyperstimulation in high risk patients. GnRH agonist triggering acts by eliciting an endogenous surge of LH and FSH, this effect on early embryonic development is still unknown. Our aim was to compare embryo morphokinetic parameters following GnRH agonist with hCG ovulation triggering, using the EmbryoScope time lapse monitoring system (TMS).

Sequential embryo scoring system can substitute for time lapse monitoring system

Time lapse scoring system has an advantage in selecting the best embryo by evaluating embryo's continuous development at certain time points. In our laboratory, we investigated if the sequential embryo-scoring system could exhibit positive effects in selecting embryos by replacing the time-lapse function with cleavage stage embryos, including early-cleavage at certain time points. All fresh, non-donor IVF cycles performed from January 2015 through August 2015 (n=485). After insemination, embryos were scored at the following 3 time points: 2-cell cleavage (26h), day 2 (40-41h), and day 3 (64-65h). The highest score of the embryo at 3 time points is as follows: 2-cell cleavage (2 points) + day 2 (5 points) + day 3 (5 points) = 12 points. In two embryos transfer, the highest total score is 24 points. The score was dependent on three factors: (number of blastomere, synchrony of cleavage, level of fragmentation). Scoring was done by subtracting scores from embryos with one or more defects. To determine a cut-off value, pregnancy rate was evaluated in one point unit from 6 points to 24 points based on two embryos transfer. Based on the cut-off value, embryos were divided into two groups, and the pregnancy and implantation rate were checked. The cut-off value was 16 points, and it was confirmed that pregnancy rate significantly improved. When the two groups (group (1) scored higher than 16 points, group (2) scored less than 15 points) based on the cut-off value were divided and compared. Patient characteristics were similar in the 2 groups. In each group, age (Group 1: 35.6±2.0 vs. Group 2: 36.0±1.9) basal AMH conc. (4.2±2.9 vs. 4.8±2.8ng/ml), and endometrial thickness (10.2±2.2 vs. 10.1±2.0mm) did not show significant differences. There were no significant differences between 2 groups in the number of MII oocytes (10.6±6.0 vs. 10.6±5.3) and number of embryo transfer (2.0±0.1 vs. 2.0±0.1). Group (1) had a significantly high pregnancy rate (68.8%, P=0.001) and implantation rate (57.1%, P=0.001) compared to that of Group (2) (41.6% and 31.6%, respectively). Our study evaluated the continuous development process of the embryo through the sequential embryo scoring system. We found that embryos with scores higher than 16 points resulted in a significantly high pregnancy rate. Therefore, the sequential embryo scoring system is effective in predicting high pregnancy rates.

The Presence of Endometrioma Does Not Impair Time-Lapse Morphokinetic Parameters and Quality of Embryos: A Study On Sibling Oocytes.

To see whether the oocytes retrieved from an ovary with an endometrioma would develop into embryos with aberrant timings of cleavage as assessed using time-lapse monitoring (TLM) and poorer morphologic quality compared with sibling oocytes from the contralateral ovary with no endometrioma in the same patient after intracytoplasmic sperm injection. This was an observational prospective study at an in vitro fertilization (IVF) center of a private hospital. It included analysis and comparison of 128 embryos (69 embryos developed from the ovary with endometrioma and 59 embryos from the contralateral ovary without endometrioma serving as controls from a total of 20 women with infertility). Morphology of the embryo was assessed twice (days 3 and 5), again by capturing images with the TLM system. Morphokinetic parameters of embryos and clinical pregnancy rates were recorded separately from ovaries with and without endometrioma and were the primary outcomes of the study. Secondary outcomes included number of retrieved oocytes, number of metaphase II (MII) oocytes, fertilization rates, and conventional morphological classification of embryos. There were no differences in terms of the following time-lapse morphokinetic parameters of embryos. The mean numbers of oocytes and MII oocytes collected from the ovary with the endometrioma were similar to those collected from the contralateral ovary without endometrioma. Fertilization rates and the percentage of embryos with top morphologic quality were also similar. According to the morphokinetic parameters, this study further strengthens the notion that removal of endometriomas before IVF is not a necessity in terms of better oocyte quality and development.

Morphokinetic analysis and embryonic prediction for blastocyst formation through an integrated time-lapse system

To describe the events associated with the blastocyst formation and implantation that occur in embryos during preimplantation development based on the largest sample size ever described with time-lapse monitoring. Observational, retrospective, single-center clinical study. University-affiliated private IVF center. A total of 7,483 zygotes from 990 first treatments of intracytoplasmic sperm injection (ICSI; 627 of oocyte donor vs. 363 autologous oocyte cycles), of which 832 blastocysts were transferred. No patient intervention. Embryos were cultured in a time-lapse monitoring system, and the embryos were transferred on day 5 after ICSI. Embryo selection was based on the multivariable model previously developed and on blastocyst morphology. Using a time-lapse system, embryo images were acquired every 15 minutes for 120 hours. Embryos cleavage time points up to the 9-cell stage (t2-t9) as well as to the morula stage (tM) and blastocyst formation (tB) were registered in hours after ICSI. Additionally, duration of the cell cycle and synchrony of the second and third cell cycles were defined. As a result, we have monitored the embryonic development of a total of 3,215 blastocysts, of which 832 were transferred. Finally, we analyzed the characteristics of embryonic development of blastocyst (phase 1) and of implanted and not implanted (phase 2) embryos as finally validated in an independent data set (phase 3). A detailed retrospective analysis of cleavage times was made for 7,483 zygotes. We analyzed 17 parameters and found several significantly correlated with subsequent blastocyst formation and implantation. The most predictive parameters for blastocyst formation were time of morula formation, tM (81.28-96.0 hours after ICSI), and t8-t5 (≤8.78 hours) or time of transition of 5-blastomere embryos to 8-blastomere embryos with a receiver operating characteristic curve (ROC) value = 0.849 (95% confidence interval [CI], 0.835-0.854; phase 1). These parameters were less predictive of implantation, with a ROC value = 0.546 (95% CI, 0.507-0.585). We also observed that time for expansion blastocyst, tEB (107.9-112.9 hours after ICSI), and t8-t5 (≤5.67 hours after ICSI) predict blastocyst implantation, with a ROC value = 0.591 (95% CI, 0.552-0.630; phase 2). The model was validated on an independent data set and gave a ROC of 0.596 (0.526-0.666; phase 3). The inclusion of kinetic parameters into score evaluation may improve blastocyst selection criteria and can predict blastocyst formation with high accuracy. We propose two multivariable models based on our findings to classify embryos according to their probabilities of blastocyst stage and implantation in the largest data set ever reported of human blastocysts.

Direct current (DC) resistivity and induced polarization (IP) monitoring of active layer dynamics at high temporal resolution

With permafrost thawing and changes in active layer dynamics induced by climate change, interactions between biogeochemical and thermal processes in the ground are of great importance. Here, active layer dynamics have been monitored using direct current (DC) resistivity and induced polarization (IP) measurements at high temporal resolution and at a relatively large scale at a heath tundra site on Disko Island on the west coast of Greenland (69°N). At the field site, the active layer is disconnected from the deeper permafrost, due to isothermal springs in the region. Borehole sediment characteristics and subsurface temperatures supplemented the DC-IP measurements. A time-lapse DC-IP monitoring system has been acquiring at least six datasets per day on a 42-electrode profile with 0.5m electrode spacing since July 2013. Remote control of the data acquisition system enables interactive adaptation of the measurement schedule, which is critically important to acquire data in the winter months, where extremely high contact resistances increase the demands on the resistivity meter. Data acquired during the freezing period of October 2013 to February 2014 clearly image the soil freezing as a strong increase in resistivity. While the freezing horizon generally moves deeper with time, some variations in the freezing depth are observed along the profile. Comparison with depth-specific soil temperature indicates an exponential relationship between resistivity and below-freezing temperature. Time-lapse inversions of the full-decay IP data indicate a decrease of normalized chargeability with freezing of the ground, which is the result of a decrease in the total unfrozen water and of the higher ion concentration in the pore-water. We conclude that DC-IP time-lapse measurements can non-intrusively and reliably image freezing patterns and their lateral variation on a 10–100m scale that is difficult to sample by point measurements. In combination with laboratory experiments, the different patterns in resistivity and chargeability changes will enable the disentanglement of processes (e.g., fluid migration and freezing, advective and diffusive heat transport) occurring during freezing of the ground. The technology can be expanded to three dimensions and also to larger scale.

Clinical validation of embryo culture and selection by morphokinetic analysis: a randomized, controlled trial of the EmbryoScope

To determine whether incubation in the integrated EmbryoScope time-lapse monitoring system (TMS) and selection supported by the use of a multivariable morphokinetic model improve reproductive outcomes in comparison with incubation in a standard incubator (SI) embryo culture and selection based exclusively on morphology. Prospective, randomized, double-blinded, controlled study. University-affiliated private invitro fertilization (IVF) clinic. Eight hundred forty-three infertile couples undergoing intracytoplasmic sperm injection (ICSI). No patient intervention; embryos cultured in SI with development evaluated only by morphology (control group) and embryos cultured in TMS with embryo selection was based on a multivariable model (study group). Rates of embryo implantation, pregnancy, ongoing pregnancy (OPR), and early pregnancy loss. Analyzing per treated cycle, the ongoing pregnancy rate was statistically significantly increased 51.4% (95% CI, 46.7-56.0) for the TMS group compared with 41.7% (95% CI, 36.9-46.5) for the SI group. For pregnancy rate, differences were not statistically significant at 61.6% (95% CI, 56.9-66.0) versus 56.3% (95% CI, 51.4-61.0). The results per transfer were similar: statistically significantdifferences in ongoing pregnancy rate of 54.5% (95% CI, 49.6-59.2) versus 45.3% (95% CI, 40.3-50.4) and not statisticallysignificantfor pregnancy rate at 65.2% (95% CI, 60.6-69.8) versus 61.1% (95% CI, 56.2-66.1). Early pregnancy loss wasstatistically significantly decreased for the TMS group with 16.6% (95% CI, 12.6-21.4) versus 25.8% (95% CI, 20.6-31.9). The implantation rate was statistically significantly increased at 44.9% (95% CI, 41.4-48.4) versus 37.1% (95% CI, 33.6-40.7). The strategy of culturing and selecting embryos in the integrated EmbryoScope time-lapse monitoring system improves reproductive outcomes. NCT01549262.

Human zygotes respond to sperm DNA damage by slowing DNA replication

OBJECTIVE: Some authors describe similar and high fertilization and pregnancy rates in non-obstructive azoospermia (NOA) patients whereas others report lower outcomes in NOA than in obstructive azoospermia (OA) patients. Sperm DNA damage is associated with reduced fertilization rates, embryo quality and pregnancy rates and higher rates of spontaneous miscarriage and childhood disease. Meanwhile, mouse zygotes respond to spermDNA damage through a non-apoptoticmechanism that acts by slowing paternal DNA replication and embryonic development. In this study, we compared the early development of embryos fertilized with NOA and OA spermatozoa by using time-lapse monitoring system. DESIGN: This is a retrospective study. MATERIALS AND METHODS: Six oocytes with ICSI using testicular spermatozoa from NOA and 35 oocytes with OA from 2013 to 2014 were used. The exact times for each embryo division and developmental parameters were calculated in hours after ICSI. Time-lapse images of each embryo were retrospectively analyzed by PrimoVision software. The developmental events observed in each embryo were at the time of the visibility of two pronuclei (2PN), syngamy, two cells and four cells. These parameters were compared between the NOA group and the OA group. RESULTS: The time of the visibility of 2PN in the NOA group (9.5 1.4 hours) was significantly earlier than that in the OA group (12.1 2.3 hours). However, the time from the visibility of 2PN to the syngamy in the NOA group (13.2 3.4 hours) was significantly longer than that in the OA group (10.6 2.7 hours). The time between the syngamy and the two cells in the NOA group (12.2 1.3 hours) was comparable with that in the OA group (16.6 10.2 hours). The time from the two cells to the four cells in the NOA group (13.5 3.5 hours) was also comparable with that in the OA group (10.0 4.2 hours). CONCLUSION: The time from the visibility of 2PN to the pronuclear fading (syngamy) after ICSI with testicular spermatozoa from NOAwas prolonged compared to that after ICSI with testicular spermatozoa from OA.

Relationships between sperm DNA fragmentation, age of donors and patients and children with psychic disorders

OBJECTIVE: Some authors describe similar and high fertilization and pregnancy rates in non-obstructive azoospermia (NOA) patients whereas others report lower outcomes in NOA than in obstructive azoospermia (OA) patients. Sperm DNA damage is associated with reduced fertilization rates, embryo quality and pregnancy rates and higher rates of spontaneous miscarriage and childhood disease. Meanwhile, mouse zygotes respond to spermDNA damage through a non-apoptoticmechanism that acts by slowing paternal DNA replication and embryonic development. In this study, we compared the early development of embryos fertilized with NOA and OA spermatozoa by using time-lapse monitoring system. DESIGN: This is a retrospective study. MATERIALS AND METHODS: Six oocytes with ICSI using testicular spermatozoa from NOA and 35 oocytes with OA from 2013 to 2014 were used. The exact times for each embryo division and developmental parameters were calculated in hours after ICSI. Time-lapse images of each embryo were retrospectively analyzed by PrimoVision software. The developmental events observed in each embryo were at the time of the visibility of two pronuclei (2PN), syngamy, two cells and four cells. These parameters were compared between the NOA group and the OA group. RESULTS: The time of the visibility of 2PN in the NOA group (9.5 1.4 hours) was significantly earlier than that in the OA group (12.1 2.3 hours). However, the time from the visibility of 2PN to the syngamy in the NOA group (13.2 3.4 hours) was significantly longer than that in the OA group (10.6 2.7 hours). The time between the syngamy and the two cells in the NOA group (12.2 1.3 hours) was comparable with that in the OA group (16.6 10.2 hours). The time from the two cells to the four cells in the NOA group (13.5 3.5 hours) was also comparable with that in the OA group (10.0 4.2 hours). CONCLUSION: The time from the visibility of 2PN to the pronuclear fading (syngamy) after ICSI with testicular spermatozoa from NOAwas prolonged compared to that after ICSI with testicular spermatozoa from OA.

Effect of sperm maturity and assisted oocyte activation on morphokinetics of early human embryos

ICSI with testicular sperm extraction (TESE) is a feasible therapeutic option for azoospermic patients, however, testicular sperm has lower fertilizability owing to its immaturity. Assisted oocyte activation (AOA) is often used to treat activation failure after ICSI. The objective of this study was to assess the effect of sperm immaturity and induced oocyte activation on early embryonic events by Time-lapse monitoring (TM). Comparative morphological assessment of ICSI zygotes. A total of 74 patients comprised of 3 groups, 57 with ejaculated sperm (EJ), 10 with AOA, and 7 with TESE. Number of zygotes in each group was 196, 48, and 65, respectively. For AOA, oocytes were briefly exposed to Ca ionophore after ICSI. Inseminated oocytes were individually cultured in a TM system (EmbryoscopeTM). Images were acquired every 10 min for up to 144 hrs. Zygotes were categorized as either those which developed to blastocysts (BL), or those which did not (non-BL). Time points of each morphokinetic event, such as 2nd polar body extrusion (PBII), pronuclear (PN) appearance/disappearance, embryonic cell divisions, and intervals between cell cleavages were analyzed. Blastulation rates for EJ, AOA, and TESE were 54.1, 41.7, and 44.6%, respectively, and all comparable. After transferring 59 EJ embryos 13 (22.0%) implanted (IMP). In EJ, the time point for the 2-cell stage was shorter in BL than in non-BL (P < 0.01), while the interval between the 2- and 3-cell stage was longer in BL (P < 0.01). Among replaced embryos, IMP zygotes developed to the 8-cell stage faster than their non-IMP (55.0 ± 6 vs. 61.1 ± 8 hrs, P < 0.01). In both AOA and TESE, time points for PN disappearance, the 2-, and 4-cell stage were shorter in BL than in non-BL (P < 0.05), while intervals between the 2- and 3-cell stage in AOA, and between the 4- and 5-cell stage in TESE were longer in BL than in non-BL (P < 0.05). Overall, in comparison to EJ zygotes, TESE extruded PBII more slowly (P < 0.05), while AOA developed faster to the 4-cell stage (P < 0.05). Faster and more synchronous cleavage was associated with higher developmental potential of zygotes irrespective of sperm origin and activation methods. Although the blastocyst formation rate was similar, sperm immaturity appeared responsible for slower first cleavage, and chemically induced activation accelerated early cleavage.

Undisturbed embryo culture in a time-lapse monitoring system improve embryo quality: a prospective cohort study

Undisturbed embryo culture in a time-lapse monitoring system improve embryo quality: a prospective cohort study

Frequency of embryo multinucleation detected by time-lapse system and its impact on pregnancy outcome

To compare the detection rate of multinucleation with the time-lapse system and conventional control timing proposed by European Society of Human Reproduction and Embryology (ESHRE) consensus and evaluate its impact on pregnancy rates. Retrospective study. A private IVF center. A total of 686 embryos from 511 intracytoplasmic sperm injection (ICSI) cycles. None. A time-lapse system was used to acquire embryo images until ET; the stored data were reviewed for the presence and persistence of multinucleation. The detection rate of multinucleation was compared with ESHRE/ALPHA consensus-proposed embryo evaluating times (23 ± 1, 26 ± 1, 44 ± 1hours). Morphokinetic characteristics of multinucleated embryos and the effect of multinucleation on pregnancy rate were researched. Multinucleation was detected in 159 embryos of 145 ICSI cycles. Using ESHRE/ALPHA consensus embryo evaluating times, only 44 (27.6%) out of 159 multinucleated embryos could be identified. In cycles with multinucleated ETs compared with cycles with no multinucleated embryos, clinical pregnancy rates (respectively, 23.4 vs. 44) and implantation rates (respectively, 23.3 vs. 43.6) were significantly lower. Time to 2-cell, 4-cell, and 6-cell stage was significantly longer in multinucleated embryos. Patient age (odds ratio [OR], 0.95; confidence interval [CI], 0.92-0.98) and presence of multinucleation (OR, 0.37; CI, 0.24-0.56) were the only significant predictors of clinical pregnancy rate. The time-lapse monitoring system seems to be a valuable tool to identify all cases with multinucleation. We conclude that the detection of multinucleation by time-lapse monitoring is associated with lower implantation and clinical pregnancy rates.

A time to look back: analysis of morphokinetic characteristics of human embryo development

To describe the times associated with the morphological changes that occur in the embryo during preimplantation development based on the largest sample size described with time lapse. Cohort study. University-affiliated private center. A total of 9,530 embryos from 1,806 intracytoplasmic sperm injection (ICSI) cycles. None. Using a time-lapse system, embryo images were acquired for at least 68 hours, in some cases reaching 120-130 hours. Embryo cleavage time points up to 8-cell-stage (t2-t8) as well as morulae (tM) and blastocyst formation (tB) were registered in hours after ICSI. Additionally, duration of the cell cycle (cc) and synchrony (s) of the second and third cell cycles were defined. Finally, four subgroups of embryos were considered: the "regular divisions" group excluded embryos with a direct cleavage from 1 to 3 or 2 to 5 cells, and the "viable 8-cell," the "viable blastocyst," and "implanted embryos" groups included only embryos viable to the 8-cell stage, blastocyst stage, or transferred and successfully implanted, respectively. Averages of times in the general population were: t2 = 27.9 hours, t3 = 38.2 hours, t4 = 40.7 hours, t5 = 51.0 hours, t6 = 54.1 hours, t7 = 56.7 hours, t8 = 59.1 hours, tM = 86.6 hours, tB = 104.1 hours, cc2 = 10.3 hours, cc3 = 12.8 hours, s2 = 2.7 hours, and s3 = 9.9 hours. Comparison between groups showed significant differences between regular divisions and viable 8 cells for t2, t3, t5, cc2, cc3, s2, and s3; between 8 cells and blastocyst for t5, t8, tM, cc3, and s2; and between blastocyst and implanted embryos for t8, tM, tB, and s2. Differences in timing related to morphology of cleavage- and blastocyst-stage embryos were detected. A time-lapse monitoring system applied to embryology allows accuracy and objectivity when defining the basis of embryo development within a clinic. The sample size is the largest ever described that provides consistent information about the normal distribution of embryo developmental timings.